Genomics ; Proteomics ; Toxicology

Objective To observe the effects of sodium arsenite (NaAsO2) on the expression of transcription factor nrf2 (Nrf2) and heme oxygenase 1 (HO-1) in Chang liver cells, and to explore the possible mechanism by using buthionine sulfoximine ( BSO), a GSH synthesis inhibitor. Methods Chang liver cells were treated with NaAsO2 at the doses of 5, 10 and 20 ?mol/L, alone, for 24 h, or pretreated with BSO (3 mmol/L, 12 h). Western blot assays were used to detect the protein expression of Nrf2 and HO-1. Results The protein expression of Nrf2 and HO-1 significantly increased in 5, 10 and 20 ?mol/L of NaAsO2 alone groups, and Nrf2 and HO-1 protein expression was up-regulated even higher by BSO pretreatment (P

OBJECTIVETo explain the essence of pungent-hot herb property express according to in vivo and in vitro studies on its effect on calmodulin on the base of the observation of the adjustment in hypothalamic-pituitary-gonad axis functions of Aconiti Lateralis Radix Praeparata, Curculiginis Rhizoma, Cinnamomi Cortex and bitter-cold herb Phellodendri Chinensis Cortex in rats under the state of yang deficiency.

METHODThe yang-deficient model was duplicated by intramuscularly injecting hydrocortisone sodium succinate powder injection. After the intervention with Aconiti Lateralis Radix Praeparata, Curculiginis Rhizoma, Cinnamomi Cortex and bitter-cold herb Phellodendri Chinensis Cortex for seven days, TSH, T3, T4, 17-OHCS, COR, T, E2 of hypothalamus-pituitary-target gland axis and other relevant indexes were detected. The calmodulin expression in livers and L02 cells cultured in vitro was detected by using Western blot.

RESULTPungent-hot herbs Aconiti Lateralis Radix Praeparata, Curculiginis Rhizoma, Cinnamomi Cortex can significantly correct indicators of hypothalamic-pituitary-gonad axis and calmodulin, whereas the bitter-cold herb Phellodendri Chinensis Cortex showed no obvious effect.

CONCLUSIONThe pungent-hot herb property expression may be closely related to calmodulin.

Animals ; Calmodulin ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Gonads ; drug effects ; metabolism ; Hypothalamo-Hypophyseal System ; drug effects ; metabolism ; Liver ; drug effects ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Yang Deficiency ; drug therapy ; metabolism

Objective To establish human choriocarcinoma JeG-3 cell line resistant to floxuridine (FUDR)and describe the characteristics of this FUDR-resistant subline.The thymidylate synthase (TS) expression level in FUDR-resistant subline was also discussed.Methods The FUDR-resistant sub-line JeG-3/FUDRA was established by intermitted exposure to grads increased FUDR.Reversed microscope was used to observe the morphological changes in FUDR-resistant sub-line.Population doubling time was calculated and compared based on the growth curve of these two cell lines,cell cycles and chromosomal ploidy were assayed with flow cytometry methods.The chemo-luminescence assay was used to detect the hormone secretion by two kinds of cell lines.The resistant index (RI) was measured by cell counting kit-8 (CCK-8)assay.Quantitative RT-PCR was used to detect the mRNA expression level of TS and we also detected the TS mRNA expression level in different doses exposed subline.Results The RI of JeG-3/FUDRA was 31.62.Compared with the JeG-3 cell,the FUDR-resistant cell line had gross changes in morphological,cell growth,cell cycles and chromosomal numbers.The ability of human chorionic gonadotrop(hCG) and progesterone secretion was lower in JeG-3/FUDRA subline.The trend of TS mRNA expression was:while exposed to low concentration of FUDR,the TS mRNA expression level was downregulated,then followed the increasing dose of the drug,the expression level of TS mRNA ascended gradually.When the terminal concentration was reached,the expression level of TS mRNA in JeG-3/FUDRA subline was higher than that of JeG-3 cell line (P＜0.05).Conclusions We established the FUDR-resistant subline of JeG-3 successfully.The TS mRNA expression level is stage-related to the different concentration and different phase in FUDR exposure.Our data suggested that TS mRNA expression level may not be used as a biomarker to predict the chemosensitivity in FUDR-based chemotherapy.

A new and effective method to produce transgenic animals was established. Without a surgical incision, the recombinant plasmid containing green fluorescence protein (GFP) cDNA was repeatedly injected into male mouse testis at multi-sites. After few weeks of the final injection, the injected male was mated with normal oestrus female to produce transgenic mice. The presence of the GFP cDNA in F1 transgenic individuals were detected by polymerase chain reaction and Southern blot hybridization, which showed that the transgenic rate of mouse F1 offspring was 41%. The transferred gene was integrated into the host genome and could be transmitted to its offspring. When the positive F1 individuals were mated with the wild type ICR mice, the F2 individuals had a transgenic rate of 37%. The results indicate that the high efficiency of gene transfer and the limited number of manipulations make the method suitable for creating a large number of transgenic animals, especially, for producing domestic animals.

Objective To detect the routine coagulation indicators in breast cancer patients receiving the trastuzumab treatment and to analyze the influence of the coagulation indicators on clinical treatment response .Methods Totally 67 hospitalized breast cancer patients receiving the trastuzumab treatment in 401 Hospital of PLA from Jan .2006 to Dec .2012 were collected and divided into the effective group(47 cases) and ineffective group(20 cases) according to the curative effect .The levels of PT ,APTT ,TT ,Fib and D‐D were statistically analyzed .The differences of coagulation indicators were compared between the two groups .The correla‐tion between the coagulation indicators with the curative effect was analyzed .Results Compared with the effective group ,PT , APTT and TT in the ineffective group were slightly extended ,while the D‐D level was slightly increased ,the differences were not statistically significant(P> 0 .05);but the Fib level was significantly increased with statistical difference(P< 0 .05) .Conclusion The breast cancer patients with ineffectiveness in the trastuzumab treatment are easily appear the high Fib status .Determining its plas‐ma concentration is conducive to judge the prognosis of the trastuzumab treatment in breast cancer patients .

The properties of Chinese materia medica are believed to be the summarization of the effects of biological performance on the various body states. Systemic discussion of chemical-factor elements, body-condition elements, biological-performance elements and their interrelationships is needed for research into the properties of Chinese materia medica. Following the practical characteristics of Chinese medicine, the three-element mathematical model was formed by introducing some mathematical concepts and methods and was used to study the cold or hot property of Chinese medicine, and to investigate the difference in biological performances of the two properties.

The theory of four properties (Qi) and five tastes (Wei) is the core of the property theory of Chinese materia medica. It is known that Qi and Wei are associated with the pharmacological effects (Xiao) of herbs. This study took records of all 365 Chinese herbs in Shennong's Classic of Materia Medica (Shennong Ben Cao Jing) as the data resource and established a three-dimensional data cube, in the purpose of finding out and analyzing the frequent patterns and valued association rules of Qi, Wei and Xiao based on Apriori algorithm. The results of this study may give rise to innovative ideas and methods in research of traditional Chinese materia medica.

ObjectiveTo study the protective effects of tert-butylhydroquinone(tBHQ) on sodium arsenite (NaAsO2)-induced cytotoxicity and oxidative injuries. Methods Chang liver cells were pretreated with tBHQ[0(control), 5, 25 μmol/L]for 24 h, and then co-treated with tBHQ(5 μmol/L) together with NaAsO2[0(control),30, 40, 50, 60 μmol/L] for another 24 h, and Alamar blue reduction rates were used to evaluate cell viability,the results were expressed as the relative ratio of Alamar blue reduction rates between the experimental group and the control group. On the other hand, Chang liver cells were pretreated with tBHQ[0(control), 5, 25 μmol/L] for24 h,and then co-treated with tBHQ(5 μmol/L) together with NaAsO2[0(control), 40, 50 μmol/L] for another 24 h,and the levels of cellular reactive oxygen species(ROS) were detected by staining cells with 2',7'-dichlorofluorescin diacetate(DCFH-DA), the results were expressed as the relative ratio of mean fluorescence intensity between the experimental group and the control group. ResultsCell viability decreased dramatically by treatment with NaAsO2(30, 40, 50, 60 μmol/L), while relieved to some extent by pretreatment with 5, 25 μmol/L tBHQ, the main effects of NaAsO2 and tBHQ, as well as their interaction were all statistically significant(F =566.57, 55.09, 14.50,all P ＜ 0.05) ; the cell viability of NaAsO2(30, 40, 50, 60 μmol/L) pretreated with tBHQ(5, 25 mol/L) were 0.75 ±0.02, 0.70 ± 0.04, 0.59 ± 0.03, 0.43 ± 0.03 and 0.75 ± 0.02, 0.73 ± 0.03, 0.65 ± 0.02, 0.50 ± 0.02, respectively,all significantly higher than corresponding NaAsO2 alone groups(0.70 ± 0.03, 0.64 ± 0.03, 0.43 ± 0.03, 0.33 ±0.01, all P ＜ 0.05), the cell viability of NaAsO2(50, 60 μmol/L) pretreated with 25 μmol/L tBHQ was higher than corresponding 5 μmol/L tBHQ pretreatment groups(all P ＜ 0.05). On the other hand, 40, 50 μmol/L of NaAsO2 significantly induced hepatocellular ROS generation, while tBHQ(5, 25 μ mol/L) pretreatment significantly decreased NaAsO2-induced intracellular ROS levels, the main effects of NaAsO2 and tBHQ, as well as their interaction were all statistically significant (F =181.78, 60.55, 4.93, all P ＜ 0.05) ; the ROS levels of NaAsO2(40, 50 μ mol/L) pretreated with tBHQ(5, 25 μmol/L) were 1.87 ± 0.09, 1.80 ± 0.07 and 1.36 ± 0.11, 1.44 ± 0.12,all significantly decreased than corresponding NaAsO2 alone groups(2.30 ± 0.18, 2.18 ± 0.17, all P ＜ 0.05),the ROS levels of NaAsO2(40, 50 μmol/L) pretreated with 25 μmol/L tBHQ decreased than corresponding 5 μmol/L tBHQ pretreatment groups (all P ＜ 0.05). ConclusiontBHQ has a certain antagonism on arsenic induced cytotoxicity and oxidative injuries.